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Abstract

Linear discriminant analysis (LDA) of single-cell fluorescence excitation
spectra (λem = 680 nm) for five species of marine phytoplankton was
used to determine whether intra-species variation among single cells precluded
discrimination among species. Single-cell spectra were recorded in an optical
trap with a custom-built spectral fluorometer. For nitrogen (N)-replete cells,
separation of all five species ( Emiliania huxleyi, a
coccolithophore,Thalassiosira pseudonana, a diatom,Dunaliella tertiolecta, a chlorophyte,Amphidinium carterae, a dinoflagellate, andRhodomonas salina, a cryptophyte) was possible using only a portion of the
excitation spectra (570-610 nm). This wavelength region gave perfect
classification of species with a minimum Fisher ratio of 62. For four species
( E. huxleyi,T. pseudonana,D. tertiolecta,andA. carterae), variations in fluorescence excitation spectra as cells were
starved of N did not impact the classification process adversely within the
chosen spectral window.R. salinacells grown with and without N showed significant differences in their
fluorescence excitation spectra but could still be classified if a different
spectral window (490-570 nm) was used. Overall, we conclude that intra-species
variation among single-cell fluorescence excitation spectra does not preclude
discrimination among species.

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